1. ISSN 1907-9850
ISOLATION AND IDENTIFICATION MOMORDICIN I FROM
LEAVES EXTRACT OF Momordica charantia L.
N. M. Puspawati
Department of Chemistry Faculty of Mathematics and Natural Sciences University of Udayana,
Bukit Jimbaran
ABSTRAK
Senyawa momordicin I telah berhasil diisolasi dari ekstrak diklorometana daun pare, Momordica charantia
L. Isolasi dilakukan dengan memfraksionasi ekstrak diklorometana yang diikuti dengan bioassay menggunakan larva
udang artemia salina, L untuk mendapatkan fraksi paling aktif. Fraksionasi dilakukan dengan kolom kromatografi
phase terbalik (C18Sep-Pak) menggunakan satu seri eluen MeOH:H2O (0-100% MeOH) secara gradien. Fraksi
paling aktif, F9 (80% MeOH : 20% H2O) selanjutnya dimurnikan dengan rekristalisasi menggunakan kloroform
sehingga dihasilkan kristal berwarna putih dengan titik leleh 125-128oC. Dari hasil interpretasi data spectra IR,
1HNMR,13CNMR, DEPT, HCOSY, HMCQC, HMBC and MS, struktur isolat aktif dielusidasi sebagai 3,7,23-
trihydroxycucurbitan-5,24-diena-19-al yang lebih dikenal dengan senyawa momordicin I.
Kata Kunci : isolasi dan identifikasii, momordicin I, Momordica charantia L., Artemia salina L.
ABSTRACT
Momordicin I has been isolated from dichloromethane extract of leaves Momordica charantia, L. The
isolation was done by fractionation the dichloromethane extract followed by bioassay using artemia salina,L larvae.
The crude extract was fractionated through reverse phase column chromatography (C18 Sep-Pak) with eluotropic
series solution of MeOH: H2O (0-100% MeOH). The most active fraction, F9 (80% MeOH-20% H2O) was purified
by recrystallisation from chloroform to yield a white crystal melted at 125-128oC. The structure of the active
compound was determined by interpretation of the spectral data (IR, 1HNMR, 13CNMR, DEPT, HCOSY, HMCOSY,
HMBC and MS) and its structure was elucidated as 3,7,23-trihydroxycucurbitan-5,24-dien-19-al which is known as
momordicin I.
Keywords : isolation and identification, momordicin I, Momordica charantia L., Artemia salina L.
INTRODUCTION (Begum, 1997). In the Philiphines, it was
reported that juice extracts from the green fruit
Momordica charantia, L is found in the of this plants are given for chronic colitis and
tropics and it is widely cultivated as vegetable bacillary dysentery; whereas the juice of the
crop. The fruits, leaves and roots of M. leaves are given in the amount of a teaspoon for
charantia have been used in Ayurweda for a children coughs. The seeds of this plant were
number of diseases, as a bitter stomachic, also reported to have potential activity as
laxative and anthelmentic (Begum, 1997). In antidiabetes (Ali, 1993), antitumor (NG, 1994),
India, the whole extract of the fruit is advocated and tripsin and elastase inhibitors (Hamato,1995)
in diseases of spleen, liver, rheumatism, gout, etc
53

2. JURNAL KIMIA 2 (1), JANUARI 2008 : 53-56
Preliminary screening for cytotoxicity The crude extract (100 mg) was
activity of some Balinese plant extracts showed dissolved in small amount of solvent and then
that dichloromethane extract of leaves filtered through Pasteur pipette. The Sep-Pak
Momordica charantia, L was toxic against column was washed with 100% MeOH and then
artemia salina larvae (Puspawati, 1997). equilibrated with water. The solvent then was
Therefore, this research was done to isolate and allowed to evaporate. Sample was injected to the
identify the active constituent of the leaves column through injector. It was then washed
Momordica charantia, L. gradiently with each 3 mL of 0, 10, 20, 30, 40,
50, 60, 70, 80, 90, 100 % MeOH respectively
with flow rate 3mL/min. Each fraction was
MATERIALS AND METHOD collected, evaporated and subjected to bioassay.
The most active fraction, F9, fraction from 80%
Materials MeOH:20% H2O was washed with
Momordica charantia, L was collected dichloromethane to give yellow solid.
from Gubug, Tabanan-Bali. The leaves parts The solid was recrystallised from
were air dried, cut up and ground to become chloroform to give a white crystal. The crystal
powder. Atemia salina, L (A.s) egg was melted at 125-128oC.
provided by technical training centre UNUD. All The active compound was characterized
solvents used in this research were proanalysis at Griffith University-Australia, using
grade. spectroscopic techniques (IR, NMR and MS).
The structure of the compound was elucidated by
Method interpretating the IR spectra, mass spectra,
1
Extraction was carried out using cold HNMR, 13 CNMR, DEPT, HCOSY, HMCOSY
solvent. Evaporation of the solvent under reduce and HMBC spectra data (Silverstein, 1991).
pressure was done on Buchi rotary evaporator.
Fractionation was performed on Waters Sep-Pak Bioassay
classic short body catridge with C18 as Bioassay was carried out using larva
stationary phase. Artemia salina L. as bioindicator to monitor
Melting point was determined on Gallen toxicity of plant extract during fractionation.
Kamp Capillary melting apparatus and The procedure was described as follows: ten
uncorrected. shrimps were transferred to each sample vial
NMR spectra were recorded on Varian containing extracts at concentration of 10, 100
Gemini operating at 400 MHz. IR spectra were and 0 µg/mL (control), using a pasteur pipette.
taken on Perkin Elmer model 1760 and mass Sea water which has been filtered was then
spectra were run on VG Flat Form 2 micromass. added to the sample vials to make 5 mL.
Bioassay using artemia salina, L (A.s) Survivors were counted after 24 hr and
larvae was conducted according to a literature percentage of death at each concentration and
procedure (Meyer, 1982). control were determined. The experiment was
done with three replicated.
Experimental
Isolation and identification RESULT AND DISCUSSION
The powder sample was extracted with
dichloromethane. The filtrate was evaporated to Fractionation and bioassay result are
give crude dichloromethane extract. summarized in Table 1.
54

3. ISSN 1907-9850
Table1. Fractionation and bioassay results
Percentage death of A.s larvae at 24
hr (%)
Frac-tion Eluen % MeOH Weigh of fraction (mg)
10
100 ppm
ppm
F1 0 - - -
F2 10 - - -
F3 20 - - -
F4 30 - - -
F5 40 - - -
F6 50 - - -
F7 60 - - -
F8 70 9.4 0 50
F9 80 10.2 90 100
F10 90 18.5 66 100
F11 100 5.4 0 20
- = not determined
The active component of leaves IR (υ max cm-1): 3418; 2945; 2871; 1708;
Momordica charantia, L was isolated as a white 1663; 1467; 1382.
crystal melted at 125-128oC. The high resolution mass spectrum of
1
HNMR (CDCl3): 9.8 (1H,19-H); 5.89 the active compound gave a molecular ion peak
(1H, 6-H); 5.08 (1H, 24-H); 4.38 (1H, 23-H); M+ = 472.6 which was consistent with the
3.89 (1H, 7-H); 3.5 (1H, 3-H); 2.4 (1H, 10-H); molecular formula C30H48O4. The IR spectrum
2.0 (1H, 8-H); 1.9 (1H, 16-H); 1.6 (1H, 16-H); (Silverstein, 1991) suggested the presence of a
1.8 (1H, 2-H); 1.7 (1H,2-H); 1.7 (3H, 27-H); 1.7 hydroxyl group (υ max cm-1= 3418), an aldehyde
(3H, 26-H); 1.7 (2H, 11-H); 1.7 (1H, 20-H); 1.6 group fuction (υ max cm-1= 1708) and C=C (υmax
(1H, 1-H); 1.5 (1H, 1-H); 1.4 (1H, 17-H); 1.2 cm-1 = 1663). The 1H- {and 13C-} spectra
(3H, 29-H); 1.2 (2H, 12-H); 1.3 (2H, 15-H); 1.0 showed the presence of an aldehyde group at δ
(3H, 30-H); 0.9 (2H, 22-H); 0.85 (3H, 21-H); 0.8 9.8-{δ 207.8}, three secondary carbinyl group at
(3H, 18-H); 0.7 (3H, 28-H). δ 4.38-{δ 66}; 3.89-{δ 65.9} and 3.5-{δ 76.5};
13
CNMR (CDCl3): 207.8 (CH, 19-C);
two trisubstituted double bond at δ 5.89-{δ 123}
145 (C, 5-C); 133 (C, 25-C); 129 (CH, 24-C);
and 5.08-{129}, and seven methyl group at δ
123(CH, 6-C); 76.5 (CH, 3-C); 66 (CH, 23-C);
0.7; 0.8; 0.85; 1.0; 1.2; 1.7 and 1.7. In addition to
65.9 (CH, 7-C); 51 (C, 9-C); 50 (CH, 17-C); 49
the above mentioned signals the DEPT spectra
(CH, 8-C); 48 (C,13-C); 46 (C, 14-C); 45 (CH2,
22-C); 42 (C, 4-C); 37 (CH, 10-C); 34 showed signals for six quartenary carbon (δ 145;
(CH2, 15-C); 33 (CH, 20-C); 30 (CH2, 12-C); ); 133; 51; 48; 46; 42), carbon of methine group (δ
29 (CH2, 16-C); 28.5 (CH2, 11-C); 28 (CH3, 29- 207.8; 129; 123; 76.5; 66; 65.9; 50; 49; 37; 33),
C); 26 (CH3, 30-C); 25 (CH3, 26-C); 23 (CH2, 1- carbon of seven methylene group (δ 45; 34; 30;
C); 21(CH2, 2-C); 18.5 (CH3, 28-C); 18 (CH3, 29; 28; 23 and 21) and carbon of seven methyl
21-C); 17 (CH3,27-C); 15 (CH3, 18-C). group (δ 18.5; 15; 18; 28; 26; 17 and 25).
MS: [M+] 472.6, corresponding to All of the information above led us to
C30H46O4. suggest that the active compound was
momordicin I or 3,7,23-trihydroxycucurbitan-5-
55

4. JURNAL KIMIA 2 (1), JANUARI 2008 : 53-56
24-dien-19-al. This compound has the same charantia on Normal and Diabetic
carbon skeleton as glycoside momordicoside Model Rats, Planta.Med,53 : 409-412
isolated from immature fruit of M. charantia Begum, S., Ahmed, S., Siddique, B. S. M Khan,
(Okabe, 1982) A., Safy, Z. S. and Arif, M, 1997,
Triterpenes, A Sterol and A Monocyclic
21 22 24 2614 Alcohol from Momordica charantia,
23
18 20 25 Phytochemistry; Elsevier Science Ltd, 44
12
11 OH
(7) : 1313-1320
19 13 17 27
Fatope, M. O., Takeda, Y., Yamashita, H.,
OHC 16
1 14 Okabe, H., and Yamauchi, T., 1990,
10
2 9
8
15 New Cucurbitane Triterpenoid from
3 28 Momordica, J. Nat. Prod, 53 (6) : 1491-
5
HO
4 7 1497
6 OH
Hamato, N., Koshiba, T., Pham, T. N., and
29 30
Tatsumi, Y., 1995, Tripsin and Elastase
Momordicin I (3,7,23,-Trihydroxycucurbitan-5,24-dien-19-al
Inhibitors From Bitter Gourd
(Momordica charantia Linn) Seeds:
The structure was confirmed by the
Purification, Amino Acid Sequences,
spectral data compared of the same compound
and Inhibitory Activities of Four New
from publication (Yasuda, 1984, Fatope, 1990).
Inhibitors, J. Biochem, 117 : 432-437
Meyer, B. N., Ferrigni, N.R and McLaughlin,
1982, Brine Shrimp: A Convenient
CONCLUSION
General Bioassay for Active Plant
Constituents, Planta Medica, 45: 31-34.
From this research, momordicin I or
NG, T. B., N., Liu, W. K., Sze, S. F., and Yeung,
3,7,23-trihydroxycucurbitan-5-24-dien-19-al was
H. W., 1994, Action of α-Momorcharin,
isolated from the leaves extract of Momordica
a Ribosome Inactivating Protein, on
charantia, L.
Cultured Tumor Cell Lines, Gen
Pharmac, 25 (1) : 75-77.
Okabe, H., Miyahara, Y., Yamauci, T., 1982,
ACKNOWLEDGEMENT
Studies on the Constituents of
Momordica charantia L
The author is very grateful to Associate
Chem.Pharm.Bull., 30 (12) : 4334-4340
Prof. David Young and Dr. Tony from Griffith
Puspawati, N. M., Tengah, I G. P., dan Mulyono,
University for the facility, support and assistance
1997, Extractions and Characterisation
given in characterization of the compound.
of Biologically Active Compounds from
Thanks also to IAEUP-DIKTI for the financial
Balinese Plants Used in Traditional
support.
Medicines, Research Report, Universitas
Udayana, Denpasar-Bali.
Silverstein, R. M., 1991, Spectrometric
REFERENCES
Identification of Organic Compound, 5th
ed, John Wiley & Sons, INC, Singapore.
Ali, L., Azad Khan, A. K., Mumun, M. I. R.,
Yasuda, M., Iwamoto, M., Okabe, H and
Mosihuzzaman, M., Nur-e-Alam, M, and
Yamauchi, T., 1984, A New Cucurbitane
Rokeya, B., 1993, Studies on
Triterpenoid From Momordica
Hypoglycemic Effects of Fruit Pulp,
charantia, Chem, Pharm, Bull, 32 (6) :
Seed, and Whole of Momordica
2044-2049
56